Final Report Summary - REGULATORYCIRCUITS (Novel Systematic Strategies for Elucidating Cellular Regulatory Circuits)
The precise regulation of gene expression has been the subject of extensive scrutiny. Nonetheless, there is a big gap between genomic characterization of transcriptional responses and predictions based on known molecular mechanisms and networks and of transcription regulation. In this project we implement an approach aimed at bridging this gap using a novel experimental strategy that exploits the recent maturation of two technologies: the use of fluorescence reporter techniques to monitor promoter activity and high-throughput genetic manipulations for the construction of combinatorial genetic perturbations. By combining these, we screen for genes that modulate the transcriptional response of target promoters, use genetic interactions between them to better resolve their functional dependencies, and build detailed quantitative models of transcriptional processes.
We use the budding yeast model organism, which allows for efficient manipulations, to dissect transcriptional responses that are prototypical of many regulatory networks in living cells. Our model is stress response to change in external environment which is mediated by multiple signaling pathways and several transcription factors. Many of these components are conserved all the way to human, although the pathways have adapted to respond to different stimuli in humans.
Our approach aims to elucidate mechanisms that are opaque to classical screens and facilitate building detailed predictive models of these responses. These results will lead to understanding of general principles that govern transcriptional networks. This is the first approach to comprehensively characterize the molecular mechanisms that modulate a transcriptional response, and arrange them in a coherent network. It will open many questions for detailed biochemical investigations, as well as set the stage to extend these ideas to use more detailed phenotypic assays and in more complex organisms.
We use the budding yeast model organism, which allows for efficient manipulations, to dissect transcriptional responses that are prototypical of many regulatory networks in living cells. Our model is stress response to change in external environment which is mediated by multiple signaling pathways and several transcription factors. Many of these components are conserved all the way to human, although the pathways have adapted to respond to different stimuli in humans.
Our approach aims to elucidate mechanisms that are opaque to classical screens and facilitate building detailed predictive models of these responses. These results will lead to understanding of general principles that govern transcriptional networks. This is the first approach to comprehensively characterize the molecular mechanisms that modulate a transcriptional response, and arrange them in a coherent network. It will open many questions for detailed biochemical investigations, as well as set the stage to extend these ideas to use more detailed phenotypic assays and in more complex organisms.
We use the budding yeast model organism, which allows for efficient manipulations, to dissect transcriptional responses that are prototypical of many regulatory networks in living cells. Our model is stress response to change in external environment which is mediated by multiple signaling pathways and several transcription factors. Many of these components are conserved all the way to human, although the pathways have adapted to respond to different stimuli in humans.
Our approach aims to elucidate mechanisms that are opaque to classical screens and facilitate building detailed predictive models of these responses. These results will lead to understanding of general principles that govern transcriptional networks. This is the first approach to comprehensively characterize the molecular mechanisms that modulate a transcriptional response, and arrange them in a coherent network. It will open many questions for detailed biochemical investigations, as well as set the stage to extend these ideas to use more detailed phenotypic assays and in more complex organisms.
We use the budding yeast model organism, which allows for efficient manipulations, to dissect transcriptional responses that are prototypical of many regulatory networks in living cells. Our model is stress response to change in external environment which is mediated by multiple signaling pathways and several transcription factors. Many of these components are conserved all the way to human, although the pathways have adapted to respond to different stimuli in humans.
Our approach aims to elucidate mechanisms that are opaque to classical screens and facilitate building detailed predictive models of these responses. These results will lead to understanding of general principles that govern transcriptional networks. This is the first approach to comprehensively characterize the molecular mechanisms that modulate a transcriptional response, and arrange them in a coherent network. It will open many questions for detailed biochemical investigations, as well as set the stage to extend these ideas to use more detailed phenotypic assays and in more complex organisms.